CONTENT

1. Introduction
2. Aim
3. Theory

4.Procedure

5.Observations

6.Conclusion

7.Reference
 INTRODUCTION
Rhizobium plural form rhizobia are prokaryotes whose main function
involves the conversion of stable nitrogen gas in the atmosphere to a
biologically useful form. Nitrogenase is an enzyme complex that reduces
dinitrogen to ammonia.

Rhizobium is a biofertilizer. Biofertilizers are substances that

contain microorganisms which when applied to the soil increase the
nutrient content and enhance the plant growth. Rhizobium,
present in the root nodules of the leguminous plants, add nitrogen
to the soil which is supplied to the plants to enhance their growth.
Rhizobium is a bacterium found in soil that helps in fixing nitrogen in
leguminous plants. It attaches to the roots of the leguminous plants and
produce nodules. These nodules fix atmospheric nitrogen and convert it
into ammonia that can be used by the plant for its growth and
development

A huge amount of energy is consumed during the nitrogen fixation and

the nitrogenase enzymes are irreversibly inactivated by oxygen.

Acetylene reduction assay is used to measure the nitrogenase activity.

Rhizobium infects the roots of leguminous plants. They are usually

found in the soil and produce nodules after infecting the roots of the
leguminous plants. As a result nitrogen gas is fixed from the atmosphere.
This nitrogen is made available to the plants that help in their growth
and development. When the legume dies there will be a breakdown of
nodules. As a result, Rhizobium is released back to the cell where it can
infect a new host.
 AIM
To study the role of Rhizobium bacteria in biofertilizer

Theory
Nitrogen-fixing bacteria, microorganisms capable of transforming
atmospheric nitrogen into fixed nitrogen (inorganic compounds usable by
plants). More than 90 percent of all nitrogen fixation is effected by these
organisms, which thus play an important role in the nitrogen cycle.

The symbiotic nitrogen-fixing bacteria invade the root hairs of host plants,
where they multiply and stimulate formation of root nodules,
enlargements of plant cells and bacteria in intimate association. Within
the nodules the bacteria convert free nitrogen to ammonia, which the host
plant utilizes for its development. To ensure sufficient nodule formation
and optimum growth of legumes (e.g., alfalfa, beans, clovers, peas,
soybeans), seeds are usually inoculated with commercial cultures of
appropriate Rhizobium species, especially in soils poor or lacking in the

required bacterium.
 MATERIALS

1. Cryema medium
2. Beaker and petridish
3. Sterile water
4. Testube
5. Ethyl alcohol
6. Scalpel
7. Glassrods
8. Forceps
 PROCEDURES

•Collecting of healthy root nodules from leguminous plants.

•washing nodules with sterile water.

•Sterlized nodules are crushed using sterile rod in a test tube containing
sterile water.

•And finally the suspension in inoculated into petriplate containing yeast

extract Mannion Congo red agar by spread plate technique.

•Incubated for 2 - 3 days at 28-30°C

 OBSERVATION
•Large gummy colonies of rhizobia will appear on YEMA plates within 4-
5 days.
 CONCLUSION

•Rhizobium, present in the root nodules of the leguminous plants,

add nitrogen to the soil which is supplied to the plants to enhance
their growth.
 REFERENCE

•https://www.microscopemaster.com/rhizobium.html